title: Diagnostic utility of isoelectric focusing and high performance liquid chromatography in neonatal cord blood screening for thalassemia and non-sickling hemoglobinopathies.
abstract: BACKGROUND
  Thalassemia syndromes are highly prevalent in Southeast Asia. In Thailand, high performance liquid chromatography (HPLC) is the most common technique routinely performed in diagnosis of thalassemia and hemoglobinopathies, while isoelectric focusing (IEF) is rarely employed. We compared the diagnostic utility of IEF and HPLC in neonatal screening for thalassemia and non-sickling hemoglobinopathies.
  
  
  METHODS
  Two-hundred and forty-one cord blood samples were analyzed using IEF and HPLC, β-thalassemia short program. The results were correlated with red cell indices and molecular analyses. Hemoglobin (Hb) Bart's was quantified only on IEF.
  
  
  RESULTS
  Of 241 newborns, IEF and HPLC yielded 85.4% and 76.4% sensitivity to identify α-thalassemia syndrome, respectively. HbBart's≥2% yielded 100% sensitivity to identify 2 α-globin gene deletions and/or mutations, while MCV≤95fl and MCH≤30pg yielded 100% sensitivity to identify 2 α-globin gene deletions. DNA analysis revealed HbE mutation in all 61 subjects with HbA2>1% on both IEF and HPLC.
  
  
  CONCLUSIONS
  IEF is an effective method in neonatal screening for thalassemia and non-sickling hemoglobinopathies. The HbBart's level, MCV and MCH are helpful for identifying α-thalassemia. The presence of HbA2 higher than 1% in cord blood indicates HbE carriers in Southeast Asian newborns.
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand; King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok 10330, Thailand. Electronic address: drnoppacharn@yahoo.com.
authors.forename: Noppacharn
authors.initials: N
authors.lastname: Uaprasert
authors.forename: Rung
authors.initials: R
authors.lastname: Settapiboon
authors.forename: Supaporn
authors.initials: S
authors.lastname: Amornsiriwat
authors.forename: Patsita
authors.initials: P
authors.lastname: Sarnthammakul
authors.forename: Tassanee
authors.initials: T
authors.lastname: Thanapat
authors.forename: Ponlapat
authors.initials: P
authors.lastname: Rojnuckarin
authors.forename: Pranee
authors.initials: P
authors.lastname: Sutcharitchan
mesh_terms: D001772:Blood Cell Count; D002851:Chromatography, High Pressure Liquid; D005312:Fetal Blood / Q000737:chemistry*; D005838:Genotype; D006453:Hemoglobinopathies / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D007231:Infant, Newborn; D007525:Isoelectric Focusing; D015997:Neonatal Screening; D013789:Thalassemia / Q000097:blood / Q000175:diagnosis* / Q000235:genetics
keywords: HPLC; IEF; Neonatal cord blood; Non-sickling hemoglobinopathies; Thalassemia